人参三七川芎提取物对复制性衰老血管平滑肌细胞骨架蛋白微丝的影响(1)
[摘要]观察人参三七川芎提取物对复制性衰老血管平滑肌细胞骨架蛋白微丝F-actin和G-actin的影响。以人主动脉平滑肌细胞(HASMC)作为研究对象，复制性衰老9代细胞作为衰老模型，实验分为青年组(5代细胞)、模型组(9代细胞)、中药低剂量组(100 mg·L-1)、中药中剂量组(200 mg·L-1)、中药高剂量组(400 mg·L-1)及白藜芦醇组(10 μmol·L-1)，药物干预时间为48 h。应用β-半乳糖苷酶特异染色法计算蓝染细胞比率，CCK-8检测细胞增殖能力，流式细胞技术分析细胞周期，免疫荧光染色观察细胞F-actin和G-actin形态的改变，Western blot 检测F-actin蛋白的表达情况。与模型组相比，中药各给药组及白藜芦醇组可以有效的降低β-半乳糖苷酶染色蓝染细胞数量，提高细胞的增殖能力，减少G0/G1期的细胞数量，同时增加S期的细胞数量，减少F-actin蛋白的表达量及应力纤维的形成，且药物干预效果明显，具有统计学意义。复制性衰老的血管平滑肌细胞可以作为衰老研究的模型，细胞骨架微丝蛋白G-actin和F-actin形态及蛋白表达在细胞衰老过程中改变明显，人参三七川芎提取物对细胞G-actin和F-actin有明显干预作用，其可能间接通过此作用延缓了血管老化的进程。
[关键词]衰老； 血管老化； 细胞骨架；人参三七川芎提取物
[Abstract]To observe the effect of extracts of ginseng， notoginseng， and Chuanxiong Rhizome on the cytoskeleton protein F-actin and G-actin of the replicative senescence vascular smooth muscle cells， with human aortic smooth muscle cells (HASMC) as the research object， and the replicative senescence 9th generation cells as the senescence models， the experiment was divided into youth group (5th generation cells)， model group (9th generation cells)， Chinese medicine low dose group (100 mg·L-1)， middle dose group (200 mg·L-1)， and high dose group (400 mg·L-1) and resveratrol group (10 μmol·L-1). The intervention time was 48 h. β-Galactosidase specific staining method was used to calculate the ratio of blue dyeing cells. CCK-8 method was used to detect the cells proliferation. The flow cytometry was used to analyze the cell cycle. Immunofluorescent staining was used to observe morphological changes of F-actin and G-actin. The western blot assay was used to determine the expression of F-actin protein. Compared with the model group， the Chinese medicine groups and resveratrol group significantly reduced the number of blue dyeing cells， improved the ability of cells proliferation， reduced the number of cells in G0/G1 phase， increased the number of cells in S phase， and reduced the protein expression of F-actin and the formation of stress fibers， with obvious intervention effect and statistically significant difference. Therefore， the replicative senescence vascular smooth muscle cells can be used as the models for senescence research， with significant changes in morphology and protein expression of cytoskeleton protein F-actin and G-actin in the process of cells aging. The extracts of ginseng， notoginseng， and Chuanxiong Rhizome have obvious intervention effect on F-actin and G-actin， and it might be indirectly associated with delaying the aging of blood vessels. (修成奎 雷燕 王强 景晓杨)
[关键词]衰老； 血管老化； 细胞骨架；人参三七川芎提取物
[Abstract]To observe the effect of extracts of ginseng， notoginseng， and Chuanxiong Rhizome on the cytoskeleton protein F-actin and G-actin of the replicative senescence vascular smooth muscle cells， with human aortic smooth muscle cells (HASMC) as the research object， and the replicative senescence 9th generation cells as the senescence models， the experiment was divided into youth group (5th generation cells)， model group (9th generation cells)， Chinese medicine low dose group (100 mg·L-1)， middle dose group (200 mg·L-1)， and high dose group (400 mg·L-1) and resveratrol group (10 μmol·L-1). The intervention time was 48 h. β-Galactosidase specific staining method was used to calculate the ratio of blue dyeing cells. CCK-8 method was used to detect the cells proliferation. The flow cytometry was used to analyze the cell cycle. Immunofluorescent staining was used to observe morphological changes of F-actin and G-actin. The western blot assay was used to determine the expression of F-actin protein. Compared with the model group， the Chinese medicine groups and resveratrol group significantly reduced the number of blue dyeing cells， improved the ability of cells proliferation， reduced the number of cells in G0/G1 phase， increased the number of cells in S phase， and reduced the protein expression of F-actin and the formation of stress fibers， with obvious intervention effect and statistically significant difference. Therefore， the replicative senescence vascular smooth muscle cells can be used as the models for senescence research， with significant changes in morphology and protein expression of cytoskeleton protein F-actin and G-actin in the process of cells aging. The extracts of ginseng， notoginseng， and Chuanxiong Rhizome have obvious intervention effect on F-actin and G-actin， and it might be indirectly associated with delaying the aging of blood vessels. (修成奎 雷燕 王强 景晓杨)